9-amino-7-(substituted)-6-demethyl-6-deoxytetracyclines

ABSTRACT

Novel 9-amino-7-(substituted amino)-6-demethyl-6-deoxytetracyclines having activity against a wide spectrum of organisms including organisms which are resistant to tetracyclines are disclosed. Also disclosed are intermediates and methods for making the novel compounds of the present invention.

This is a divisional of co-pending application Ser. No. 07/771,697,filed on Oct. 04, 1991, now U.S. Pat. No. 5,281,628.

FIELD OF THE INVENTION

The invention relates to novel[4S-(4α,12aα)]-9-amino-4-(dimethylamino)-7-(substitutedamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamides, hereinafter called9-amino-7-(substituted amino)-6-demethyl-6-deoxytetracyclines, whichexhibit antibiotic activity against a wide spectrum of organismsincluding organisms which are resistant to tetracyclines and are usefulas antibacterial agents. The invention also relates to novel7-(substituted amino)-9-nitro-6-demethyl-6-deoxytetracycline compoundsuseful for making the novel compounds of the present invention and tonovel methods for producing the novel compounds and intermediatecompounds.

DESCRIPTION OF THE PRIOR ART

A variety of tetracycline antibiotics have been synthesized anddescribed for the treatment of infectious diseases in man and animalssince 1947. Tetracyclines inhibit protein synthesis by binding to the30S subunit of the bacterial ribosome preventing binding of aminoacylRNA (Chopra, Handbook of Experimental Pharmacology, Vol. 78, 317-392,Springer-Verlag, 1985). Resistance to tetracyclines has emerged amongmany clinically important microorganisms which limit the utility ofthese antibiotics. There are two major mechanisms of bacterialresistance to tetracyclines: a) energy-dependent efflux of theantibiotic mediated by proteins located in the cytoplasmic membranewhich prevents intracellular accumulation of tetracycline (S.B. Levy, etal., Antimicrob. Agents Chemotherapy 33, 1373-1374 (1989); and b)ribosomal protection mediated by a cytoplasmic protein which interactswith the ribosome such that tetracycline no longer binds or inhibitsprotein synthesis (A.A. Salyers, B.S. Speers and N.B. Shoemaker, Mol.Microbiol, 4:151-156, 1990). The efflux mechanism of resistance isencoded by resistance determinants designated tetA-tetL. They are commonin many Gram-negative bacteria (resistance genes Class A-E), such asEnterobacteriaceae, Pseudomonas, Haemoohilus and Aeromonas, and inGram-positive bacteria (resistance genes Class K and L), such asStaphylococcus, Bacillus and Streotococcus. The ribosomal protectionmechanism of resistance is encoded by resistance determinants designatedTetM, N and O, and is common in Staphylococcus, Streptococcus,Camoylobacter, Gardnerella, Haemophilus and Mycoplasma (A.A. Salyers,B.S. Speers and N.B. Shoemaker, Mol. Microbiol, 4:151-156 1990).

A particularly useful tetracycline compound is7-(dimethylamino)-6-demethyl-6-deoxytetracycline, known as minocycline(see U.S. Pat. No. 3,148,212, RE 26,253 and 3,226,436 discussed below).However, strains harboring the tetB (efflux in gram-negative bacteria)mechanism, but not tetK (efflux in Staphylococcus) are resistant tominocycline. Also, strains carrying tetM (ribosomal protection) areresistant to minocycline. This invention describes the synthesis ofnovel tetracycline compounds which demonstrate significant in vitro andin vivo activity vs. tetracycline and minocycline susceptible strainsand some tetracycline and minocycline resistant strains, that is, thoseharboring the tetM (ribosomal protection) resistance determinants.

Duggar, U.S. Pat. No. 2,482,055, discloses the preparation ofAureomycin® (I) by fermentation which have antibacterial activity.Growich et al., U.S. Pat. No. 3,007,965, disclose improvements to thefermentation preparation of I. Neither of these patents teaches orsuggests the 6-demethyl-6-deoxytetracyclines. ##STR1## Beereboom et al.,U.S. Pat. No. 3,043,875 discloses tetracycline derivatives of theformulae (II) and (III) where R is H or CH₃ ; R₁ is H and when R is CH₃,OH; R₂ is H and N(CH₃)₂ ; X and Y are halogen; Z is H and halogen and Bis bromo, chloro and iodo, which have antibacterial activity. Thispatent does not teach or suggest the inclusion of both di(loweralkyl)amino or mono(lower alkyl)amino substituents (at Y or Z) and anamino function (at B). ##STR2## Boothe et al., U.S. Pat. No. 3,148,212,reissued as RE26,253, and Petisi et al., U.S. Pat. No. 3,226,436,discloses tetracycline derivatives of the formula (IV) wherein R ishydrogen or methyl and R₁ and R₂ is hydrogen, mono(lower alkyl)amino ordi(lower alkyl)amino with the proviso that R₁ and R₂ cannot both behydrogen, which are useful for treating bacterial infections. Thispatent does not teach or suggest the inclusion of a 9-aminofunctionality (at R₂). ##STR3## Blackwood et al., U.S. Pat. No.3,200,149 discloses tetracycline derivatives of the formulae (V) and(VI) and reduction products thereof wherein Y may be hydrogen orhydroxyl, X may be hydrogen, chloro, iodo, or bromo, X₁ may be hydrogen,amino, and lower alkanoylamino, X₂ may be hydrogen or nitro and Z ischloro or fluoro which possess microbiological activity. This patentdoes not teach or suggest the inclusion of both a di(lower alkyl)aminogroup (at X) and another nitrogen functionality (at X₁) on the6-demethyl-6-deoxytetracycline nucleus. ##STR4## Petisi et al., U.S.Pat. No. 3,338,963 discloses tetracycline compounds of the formula (VII)wherein R₁ and R₂ are hydrogen, nitro, amino, formylamino, acetylamino,p-(dihydroxyboryl)benzoylamino, p-(aminobenzenesulfonyl)amino, chlorine,bromine or diazonium with the proviso that R₁ and R₂ may not both behydrogen and with the further proviso that when R₁ is chlorine orbromine, R₂ may not be hydrogen and vice versa, R₃ is hydrogen or methyland R₄ is hydrogen or hydroxy, which have broad-spectrum antibacterialactivity. This patent does not teach or suggest the inclusion of bothdi(lower alkyl)amino or mono(lower alkyl)amino substituents (at R₁) andamino substituents (at R₂). ##STR5## Bitha et al., U.S. Pat. No.3,341,585 discloses tetracycline compounds of the formula (VIII) whereinR₅ is hydrogen, α-hydroxy or β-hydroxy R₆ is α-methyl or β-methyl, andR₇ and R₉ are each hydrogen, mono(lower alkyl)amino or di(loweralkyl)amino with the proviso that R₇ and R₉ cannot both be hydrogen andwith the further proviso that when R₅ is hydrogen then R₆ is α-methyl. Apreferred embodiment of the general formula (VIII) is when R₅ isα-hydroxy or β-hydrox R₆ is α-methyl or β-methyl, R₇ is di(loweralkyl)amino and R₉ is hydrogen, which have broad-spectrum antibacterialactivity. This patent does not teach or suggest the inclusion of bothdi(lower alkyl)amino or mono(lower alkyl)amino substituents (at R₇) andamino substituents (at R₉). ##STR6## Shu, U.S. Pat. No. 3,360,557discloses 9-hydroxytetracyclines of the formula (IX) wherein R₁ ishydrogen or hydroxy, R₂ is hydrogen or hydroxy, R₃ is hydrogen ormethyl, R₂ and R₃ taken together is methylene, and R₄ is hydrogen,halogen, nitro, amino, mono(lower alkyl)amino or di(lower alkyl)amino,which have been found to possess antibacterial activity. This patent isrestricted to 9-hydroxytetracyclines and does not teach or suggest thepresently claimed compounds. ##STR7## Zambrano, U.S. Pat. No. 3,360,561discloses a process for preparing 9-nitrotetracyclines of the formula(X) wherein R₅ is hydrogen or hydroxy, R₁ is hydrogen or hydroxy, R₆ ishydrogen or methyl, R₁ and R₆ taken together is methylene, R₇ ishydrogen, chloro or nitro and R₉ is hydrogen or nitro with the provisothat R₇ and R₉ cannot both be hydrogen. This patent does not teach orsuggest the inclusion of both a di(lower alkyl)amino or mono(loweralkyl)amino substituent (at R₇) and an amino functionality (at R₉).##STR8## Martell et al., U.S. Pat. No. 3,518,306 discloses 7-and/or9-(N-nitrosoalkylamino)-6-demethyl-6-deoxytetracyclines of the formula(XI) which possess in vivo antibacterial activity. This patent does notteach or suggest the inclusion of both a di(lower alkyl)amino ormono(lower alkyl)amino substituent (at C-7) and an amino functionality(at C-9) ##STR9##

In U.S. Pat. No. 5,021,407, a method of overcoming the resistance oftetracycline resistant bacteria is disclosed. The method involvesutilizing a blocking agent compound in conjunction with a tetracyclinetype antibiotic. This patent does not disclose novel tetracyclinecompounds which themselves have activity against resistant organisms.

In summary, none of the above patents teach or suggest the novelcompounds of this application. In addition, none of the above patentsteach or suggest novel tetracycline compounds having activity againsttetracycline and minocycline resistant strains as well as strains whichare normally susceptible to tetracyclines.

SUMMARY OF THE INVENTION

This invention is concerned with novel 9-amino-7-(substitutedamino)-6-demethyl-6-deoxytetracyclines, represented by formula I and II,which have antibacterial activity, with methods of treating infectiousdiseases in warm-blooded animals employing these new compounds; withmethods of treating or controlling veterinary diseases; withpharmaceutical preparations containing these compounds; with novelintermediate compounds and processes for the production of thesecompounds. More particularly, this invention is concerned with compoundsof formula I which have enhanced in vitro and in vivo antibacterialactivity against tetracycline resistant strains as well as a high levelof activity against strains which are normally susceptible totetracyclines. ##STR10##

In formula I and II, R=NR₁ R₂,

and when R₁ =hydrogen, R₂ =methyl, ethyl, n-propyl, 1-methylethyl,n-butyl, 1-methylpropyl, 2-methylpropyl or 1,1-dimethylethyl;

and when R₁ =methyl or ethyl, R₂ =methyl, ethyl, n-propyl,1-methylethyl, n-butyl, 1-methylpropyl or 2-methylpropyl;

and when R₁ =n-propyl, R₂ =n-propyl, 1-methylethyl, n-butyl,1-methylpropyl or 2-methylpropyl;

and when R₁ =1-methylethyl, R₂ =n-butyl, 1-methylpropyl or2-methylpropyl;

and when R₁ =n-butyl, R₂ =n-butyl, 1-methylpropyl or 2-methylpropyl;

and when R₁ =1-methylpropyl, R₂ =2-methylpropyl;

R₃ is selected from hydrogen, straight or branched (C₁ -C₃)alkyl groupsuch as methyl, ethyl, n-propyl or 1-methylethyl; (C₆ -C₁₀)aryl groupsuch as phenyl, α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such asbenzyl, 1-phenylethyl, 2-phenylethyl or phenylpropyl; a heterocyclegroup such as 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁-C₃)alkyl substituted pyridyl, benzofuranyl, benzothienyl, quinolinylor--(CH₂)_(n) COOR₅ when n=1-4and R₅ is selected from hydrogen; straightor branched (C₁ -C₃)alkyl group such as methyl, ethyl, n-propyl or1-methylethyl; or (C₆ -C₁₀) aryl group such as phenyl, α-naphthyl,β-naphthyl; R₄ is selected from hydrogen; straight or branched (C₁-C₃)alkyl group such as methyl, ethyl, n-propyl or 1-methylethyl; C₆-C₁₀)aryl group such as phenyl, α-naphthyl or β-naphthyl; (C₇-C₉)aralkyl group such as benzyl, 1-phenylethyl, 2-phenylethyl orphenylpropyl; a heterocycle group such as 2 or 3-furanyl, 2 or3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkyl substituted pyridyl,benzofuranyl, benzothienyl, quinolinyl or--(CH₂)_(n) COOR₆ when n=1-4and R₆ is selected from hydrogen; straight or branched (C₁ -C₃)alkylsuch as methyl, ethyl, n-propyl or 1-methylethyl; or (C₆ -C₁₀)aryl suchas phenyl, α-naphthyl or β-naphthyl; or R₃ and R₄ taken together are--(CH₂)₂ W(CH₂)--₂, wherein W is selected from (CH₂)_(n) and n=0-1,--NH, --N(C₁ -C₃)alkyl, --N(C₁ -C₄)alkoxy, oxygen, sulfur or substitutedcongeners selected from (L or D) proline, ethyl (L or D) prolinate,morpholine, pyrrolidine or piperidine; and the pharmacologicallyacceptable organic and inorganic salts, and metal complexes.

Particularly preferred are compounds according to the above formula Iand II in which R=NR₁ R₂,

and when R₁ =hydrogen, R₂ =ethyl, n-propyl, 1-methylethyl, n-butyl,1-methylpropyl, 2-methylpropyl or 1,1-dimethylethyl;

and when R₁ =methyl, R₂ =methyl, ethyl, n-propyl, n-butyl,1-methylpropyl or 2-methylpropyl;

and when R₁ =ethyl, R₂ =ethyl, n-propyl, n-butyl or 2-methylpropyl;

and when R₁ =n-propyl, R₂ =n-propyl, n-butyl or 2-methylpropyl;

and when R₁ =n-butyl, R₂ =n-butyl or 2-methylpropyl;

R₃ is selected from hydrogen, straight or branched (C₁ -C₃)alkyl groupsuch as methyl, ethyl, n-propyl or 1-methylethyl; (C₆ -C₁₀)aryl groupsuch as phenyl, α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such asbenzyl, 1-phenylethyl, 2-phenylethyl or phenylpropyl; a heterocyclegroup such as 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁-C₃)alkyl substituted pyridyl, benzofuranyl, benzothienyl, quinolinylor--(CH₂)_(n) COOR₅ when n=1-4 and R₅ is selected from hydrogen;straight or branched (C₁ -C₃)alkyl group such as methyl, ethyl, n-propylor 1-methylethyl; or (C₆ -C₁₀) aryl group such as phenyl, α-naphthyl,β-naphthyl; R₄ is selected from hydrogen; straight or branched (C₁-C₃)alkyl group such as methyl, ethyl, n-propyl or 1-methylethyl; (C₆-C₁₀)aryl group such as phenyl, α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such as benzyl, 1-phenylethyl, 2-phenylethyl orphenylpropyl; a heterocycle group such as 2 or 3-furanyl, 2 or3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkyl substituted pyridyl,benzofuranyl, benzothienyl, quinolinyl or--(CH₂)_(n) COOR₆ when n=1-4and R₆ is selected from hydrogen; straight or branched (C₁ -C₃)alkylsuch as methyl, ethyl, n-propyl or 1-methylethyl; or (C₆ -C₁₀)aryl suchas phenyl, α-naphthyl or β-naphthyl; or R₃ and R₄ taken together are--(CH₂)₂ W(CH₂)--₂, wherein W is selected from (CH₂)_(n) and n=0-1,--NH, --N(C₁ -C₃)alkyl, --N(C₁ -C₄)alkoxy, oxygen, sulfur or substitutedcongeners selected from (L or D) proline, ethyl (L or D) prolinate,morpholine, pyrrolidine or piperidine; and the pharmacologicallyacceptable organic and inorganic salts, and metal complexes.

Most particularly preferred are compounds according to the above formulaI and II in which R=NR₁ R₂,

and when R₁ =hydrogen, R₂ =ethyl, n-propyl or 1-methylethyl;

and when R₁ =methyl, R₂ =methyl, ethyl or n-propyl;

and when R₁ =ethyl, R₂ =ethyl;

R₃ is selected from hydrogen, straight or branched (C₁ -C₃) alkyl groupsuch as methyl, ethyl, n-propyl or 1-methylethyl; (C₆ -C₁₀)aryl groupsuch as phenyl, α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such asbenzyl, 1-phenylethyl, 2-phenylethyl or phenylpropyl; a heterocyclegroup such as 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁-C₃)alkyl substituted pyridyl, benzofuranyl, benzothienyl, quinolinylor--(CH₂)_(n) COOR₅ when n=1-4 and R₅ is selected from hydrogen;straight or branched (C₁ -C₃)alkyl group such as methyl, ethyl, n-propylor 1-methylethyl; or (C₆ -C₁₀) aryl group such as phenyl, α-naphthyl,β-naphthyl; R₄ is selected from hydrogen; straight or branched (C₁-C₃)alkyl group such as methyl, ethyl, n-propyl or 1-methylethyl; (C₆-C₁₀)aryl group such as phenyl, α-naphthyl or β-naphthyl; (C₇ -C.sub.9)aralkyl group such as benzyl, 1-phenylethyl, 2-phenylethyl orphenylpropyl; a heterocycle group such as 2 or 3-furanyl, 2 or3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkyl substituted pyridyl,benzofuranyl, benzothienyl, quinolinyl or--(CH₂)_(n) COOR₆ when n=1-4and R₆ is selected from hydrogen; straight or branched (C₁ -C₃)alkylsuch as methyl, ethyl, n-propyl or 1-methylethyl; or (C₆ -C₁₀)aryl suchas phenyl, α-naphthyl or β-naphthyl; or R₃ and R₄ taken together are--(CH₂)₂ W(CH₂)--₂, wherein W is selected from (CH₂)_(n) and n=0-1,--NH, --N(C₁ -C₃)alkyl, --N(C₁ -C₄)alkoxy, oxygen, sulfur or substitutedcongeners selected from (L or D) proline, ethyl (L or D) prolinate,morpholine, pyrrolidine or piperidine; and the pharmacologicallyacceptable organic and inorganic salts, and metal complexes.

Also included in the present invention are compounds useful asintermediates for producing the above compounds of formula I. Suchintermediate compounds include those having the formulae: ##STR11##wherein: R=NR₁ R₂,

and when R₁ =hydrogen, R₂ =methyl, ethyl, n-propyl, 1-methylethyl,n-butyl, 1-methylpropyl, 2-methylpropyl or 1,1-dimethylethyl;

and when R₁ =methyl or ethyl, R₂ =methyl, ethyl, n-propyl,1-methylethyl, n-butyl, 1-methylpropyl or 2-methylpropyl;

and when R₁ =n-propyl, R₂ =n-propyl, 1-methylethyl, n-butyl,1-methylpropyl or 2-methylpropyl;

and when R₁ =1-methylethyl, R₂ =n-butyl, 1-methylpropyl or2-methylpropyl;

and when R₁ =n-butyl, R₂ =n-butyl, 1-methylpropyl or 2-methylpropyl;

and when R₁ =1-methylpropyl, R₂ =2-methylpropyl;

R₃ is selected from hydrogen, straight or branched (C₁ -C₃)alkyl groupsuch as methyl, ethyl, n-propyl or 1-methylethyl; (C₆ -C₁₀)aryl groupsuch as phenyl, α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such asbenzyl, 1-phenylethyl, 2-phenylethyl or phenylpropyl; a heterocyclegroup such as 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁-C₃)alkyl substituted pyridyl, benzofuranyl, benzothienyl, quinolinylor--(CH₂)_(n) COOR₅ when n=1-4 and R₅ is selected from hydrogen;straight or branched (C₁ -C₃)alkyl group such as methyl, ethyl, n-propylor 1-methylethyl; or (C₆ -C₁₀) aryl group such as phenyl, α-naphthyl,β-naphthyl; R₄ is selected from hydrogen; straight or branched (C₁-C₃)alkyl group such as methyl, ethyl, n-propyl or 1-methylethyl; (C₆-C₁₀)aryl group such as phenyl, α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such as benzyl, 1-phenylethyl, 2-phenylethyl orphenylpropyl; a heterocycle group such as 2 or 3-furanyl, 2 or3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkyl substituted pyridyl,benzofuranyl, benzothienyl, quinolinyl or--(CH₂)_(n) COOR₆ when n=1-4and R₆ is selected from hydrogen; straight or branched (C₁ -C₃)alkylsuch as methyl, ethyl, n-propyl or 1-methylethyl; or (C₆ -C₁₀)aryl suchas phenyl, α-naphthyl or β-naphthyl; or R₃ and R₄ taken together are--(CH₂)₂ W(CH₂)--₂, wherein W is selected from (CH₂)_(n) and n=0-1,--NH, --N(C₁ -C₃)alkyl, --N(C₁ -C₄)alkoxy, oxygen, sulfur or substitutedcongeners selected from (L or D) proline, ethyl (L or D) prolinate,morpholine, pyrrolidine or piperidine; and the pharmacologicallyacceptable organic and inorganic salts, and metal complexes.

Particularly preferred are compounds according to the above formula IIIand IV in which R=NR₁ R₂,

and when R₁ =hydrogen, R₂ =ethyl, n-propyl, 1-methylethyl, n-butyl,1-methylpropyl, 2-methylpropyl or 1,1-dimethylethyl;

and when R₁ =methyl, R₂ =methyl, ethyl, n-propyl, n-butyl,1-methylpropyl or 2-methylpropyl;

and when R₁ =ethyl, R₂ =ethyl, n-propyl, n-butyl or 2-methylpropyl;

and when R₁ =n-propyl, R₂ =n-propyl, n-butyl or 2-methylpropyl;

and when R₁ =n-butyl, R₂ =n-butyl or 2-methylpropyl;

R₃ is selected from hydrogen, straight or branched (C₁ -C₃)alkyl groupsuch as methyl, ethyl, n-propyl or 1-methylethyl; (C₆ -C₁₀)aryl groupsuch as phenyl, α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such asbenzyl, 1-phenylethyl, 2-phenylethyl or phenylpropyl; a heterocyclegroup such as 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁-C₃)alkyl substituted pyridyl, benzofuranyl, benzothienyl, quinolinylor--(CH₂)_(n) COOR₅ when n=1-4 and R₅ is selected from hydrogen;straight or branched (C₁ -C₃)alkyl group such as methyl, ethyl, n-propylor 1-methylethyl; or (C₆ -C₁₀) aryl group such as phenyl, α-naphthyl,β-naphthyl; R₄ is selected from hydrogen; straight or branched (C₁-C₃)alkyl group such as methyl, ethyl, n-propyl or 1-methylethyl: (C₆-C₁₀)aryl group such as phenyl, α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such as benzyl, 1-phenylethyl, 2-phenylethyl orphenylpropyl: a heterocycle group such as 2 or 3-furanyl, 2 or3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkyl substituted pyridyl,benzofuranyl, benzothienyl, quinolinyl or--(CH₂)_(n) COOR₆ when n=1-4and R₆ is selected from hydrogen; straight or branched (C₁ -C₃)alkylsuch as methyl, ethyl, n-propyl or 1-methylethyl; or (C₆ -C₁₀)aryl suchas phenyl, α-naphthyl or β-naphthyl; or R₃ and R₄ taken together are--(CH₂)₂ W(CH₂)--₂, wherein W is selected from (CH₂)_(n) and n=0-1,--NH, --N(C₁ -C₃)alkyl --N(C₁ -C₄)alkoxy, oxygen, sulfur or substitutedcongeners selected from (L or D) proline, ethyl (L or D) prolinate,morpholine, pyrrolidine or piperidine; and the pharmacologicallyacceptable organic and inorganic salts, and metal complexes.

Most particularly preferred are compounds according to the above formulaIII and IV in which R=NR₁, R₂, and when R₁ =hydrogen, R₂ =ethyl,n-propyl or 1-methylethyl;

and when R₁ =methyl, R₂ =methyl, ethyl or n-propyl;

and when R₁ =ethyl, R₂ =ethyl;

R₃ is selected from hydrogen, straight or branched (C₁ -C₃)alkyl groupsuch as methyl, ethyl, n-propyl or 1-methylethyl; (C₆ -C₁₀)aryl groupsuch as phenyl, α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such asbenzyl, 1-phenylethyl, 2-phenylethyl or phenylpropyl; a heterocyclegroup such as 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁-C₃)alkyl substituted pyridyl, benzofuranyl, benzothienyl, quinolinylor--(CH₂)_(n) COOR₅ when n=1-4 and R₅ is selected from hydrogen;straight or branched (C₁ -C₃)alkyl group such as methyl, ethyl, n-propylor 1-methylethyl; or (C₆ -C₁₀) aryl group such as phenyl, α-naphthyl,β-naphthyl; R₄ is selected from hydrogen; straight or branched (C₁-C₃)alkyl group such as methyl, ethyl, n-propyl or 1-methylethyl; (C₆-C₁₀)aryl group such as phenyl, α-naphthyl or β-naphthyl; (C₇ -C₉)aralkyl group such as benzyl, 1-phenylethyl, 2-phenylethyl orphenylpropyl; a heterocycle group such as 2 or 3-furanyl, 2 or3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkyl substituted pyridyl,benzofuranyl, benzothienyl, quinolinyl or--(CH₂)_(n) COOR₆ when n=1-4and R₆ is selected from hydrogen; straight or branched (C₁ -C₃)alkylsuch as methyl, ethyl, n-propyl or 1-methylethyl; or (C₆ -C₁₀)aryl suchas phenyl, α-naphthyl or β-naphthyl; or R₃ and R₄ taken together are--(CH₂)₂ W(CH₂)₋₋ ₂, wherein W is selected from (CH₂)_(n) and n=0-1,--NH, --N(C₁ -C₃)-alkyl, --N(C₁ -C₄)alkoxy, oxygen, sulfur orsubstituted congeners selected from (L or D) proline, ethyl (L or D)prolinate, morpholine, pyrrolidine or piperidine; and thepharmacologically acceptable organic and inorganic salts, and metalcomplexes.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The novel compounds of the present invention may be readily prepared inaccordance with the following schemes.

The starting 7-(substituted amino)-6-demethyl-6-deoxytetracyclinesdescribed in formula 1, wherein R=NR₁ R₂ and R₁ =R₂ (1a) and R=NHR₂ (1b)or the salts thereof are prepared by procedures known to those skilledin the art including those described in U.S. Pat. Nos. 3,226,436 and3,518,306. ##STR12##

The starting 7-(substituted amino)-6-demethyl-6-deoxytetracyclinesdescribed in formula 1 wherein R=NR₁ R₂ and R₁ ≠R₂ (1c) are preparedaccording

to Scheme 1. ##STR13## In accordance with Scheme I, a7-(monoalkylamino)-6-demethyl-6-doxytetracycline, 1b, in which R=NHR₂,is reductively alkylated with an aldehyde to give an unsymmetricaldialkylamino, 1c. ##STR14##

In accordance with Scheme II, a 7-(substituted amino)-6-demethyl6-deoxytetracycline or its salts, 1a-1c, is treated with

a) a metal nitrate salt; such as calcium, potassium or sodium; and astrong acid; such as sulfuric acid, trifluoroacetic acid,methanesulfonic acid or perchloric acid or

b) nitric acid and a strong acid; such as sulfuric acid, trifluoroaceticacid, methanesulfonic acid or perchloric acid; to form the corresponding7-(substituted amino)-9-nitro-6-demethyl-6-deoxytetracycline 2.

To produce the 9-(amino)-7-(substitutedmino)-6-demethyl-6-deoxytetracyclines of the present invention, compound2 or its salts is treated with hydrogen in an acid alcohol solvent,preferably 2-methoxyethanol, in the presence of a suitable catalyst suchas, for example:

a) any supported catalyst; such as 0.5-25% palladium-on-carbon, 0.5-25%palladium-on-barium sulfate, 0.5-25% platinum-on-carbon or 0.5-25%rhodium-on-carbon;

b) any reducible metal oxide catalyst; such as Raney nickel or platinumoxide; or

c) a homogeneous hydrogenation catalyst; such astris(triphenylphosphine)rhodium (I) chloride; to obtain the9-amino-7-(substituted amino)-6-demethyl-6-deoxytetracycline, 3.

Alternatively, the 9-(amino)-7-(substitutedamino)-6-demethyl-6-deoxytetracyclines of the present invention areobtained by treating with:

a) stannous chloride dihydrate as described by R.B. Wordward, Org. Syn.,Coll. Vol. 3, 453 (1955);

b) a soluble metal sulfide, preferably sodium sulfide, in alcoholicsolvents as described by G.R. Robertson, Org. Syn., Coll. Vol. 1, 52(1941);

c) an active metal in mineral acid; such as iron, tin or zinc in dilutehydrochloric acid;

d) active metal couples; such as copper-zinc, tin-mercury or aluminumamalgam in dilute acid; or

e) transfer hydrogenation using triethylammonium formate and a supportedcatalyst as described by I.D. Entwistle et al., J. Chem. Soc., Perkin 1,443 (1977). ##STR15##

In accordance with Scheme III, Z=NO₂ or NH₂ ; compound 4 is selectivelyN-alkylated in the presence of formaldehyde and either a primary aminesuch as methylamine, ethylamine, benzylamine, methyl glycinate, (L or D)lysine, (L or D) alanine or their substituted congeners; or a secondaryamine such as morpholine, pyrrolidine, piperidine or their substitutedcongeners to give the corresponding Mannich base adducts, 5. of thebiologically active 7-(substitutedamino)-6-demethyl-6-deoxytetracyclines. Contemplated equivalents includethose substituted morpholine, pyrrolidine or piperidine moieties whereinthe substituents are chosen to provide the requisite increase insolubility without adversely affecting antibacterial activity.

The 9-amino-7-(substituted amino)-6-demethyl-6-deoxytetracyclines, 3,may also be obtained as metal complexes such as aluminum, calcium, iron,magnesium, manganese and complex salts; inorganic and organic salts andcorresponding Mannich base adducts using methods known to those skilledin the art. Preferably, the 9-amino-7-(substitutedamino)-6-demethyl-6-deoxytetracyclines, are obtained as inorganic saltssuch as hydrochloric, hydrobromic, hydroiodic, phosphoric, nitric orsulfate; or organic salts such as acetate, benzoate, citrate, cysteineor other amino acids, fumarate, glycolate, maleate, succinate, tartrate,alkylsulfonate or arylsulfonate. In all cases, the salt formation occurswith the C(4)-dimethylamino group. The salts are preferred for oral andparenteral administration.

Biological Activity Methods for In Vitro Antibacterial Evaluation (Table1)

The minimum inhibitory concentration (MIC), the lowest concentration ofthe antibiotic which inhibits growth of the test organism, is determinedby the microtiter broth dilution method using 0.1 ml Muller-Hinton IIbroth (Baltimore Biological Laboratories) per well. A suitable oxygenscavenger (i.e., cysteine or dithiothreitol) is added to the assaymedium for the testing of compounds according to Formula I because ofthe sensitivity of those compounds to oxidation. An inoculum level of1-5×10⁵ CFU/ml and a range of antibiotic concentrations (32 -0.004μg/ml) are used. MIC's were determined after the plates were incubatedfor 18 hours at 35° C. in a forced air incubator.

e. Coli In Vitro Protein Translation System (Table 2)

The E. coli in vitro translation system can be used to study not onlythe mechanism of protein translation itself, but also the effect thatvarious compounds may have on protein synthesis. The system can be setup to function as a coupled transcription and translation system or as atranslation only system depending on whether DNA or RNA is added toinitiate protein synthesis. In this way, compounds affecting either RNAsynthesis and/or protein synthesis can be studied. Protein synthesis ismonitored by the incorporation of radiolabeled amino acids intotrichloroacetic acid precipitable material. The system used is basedupon literature methods [G. Zubay, Ann. Rev. Genet., 7: 267-287(1973)and J. Collins, Gene, 6: 28-42 (1979)].

The system used to study tetracycline protein synthesis inhibition is asfollows:

An S30 extract (supernatant from a 30,000×G centrifugation of lysedcells) of either tetracycline sensitive or tetracycline resistant(tetM+) cells is combined with a mixture of low molecular weightcompounds required for protein synthesis which include a mixture of 19amino acids, methionine, ³⁵ S-methionine, plasmid template DNA andeither dimethylsulfoxide (DMSO) or the tetracycline to be testeddissolved and diluted in DMSO. This mixture is incubated at 37° C. for30 minutes. Following the incubation, 2.5 μl of the 10 μl reaction isremoved and added to 0.5 ml of 1N sodium hydroxide. The solution isincubated an additional 15 minutes at 37 ° C., to destroy any m-RNA andt-RNA. The incorporation of ³⁵ S-methionine is determined byprecipitating the high molecular weight material in the sodium hydroxidealiquot with trichloroacetic acid (TCA), collecting the precipitatedmaterial on Whatman G/FC filters, drying the filters and counting theradioactivity retained on the filter. Percent inhibition (P.I.) ofprotein synthesis is determined by the following equation: ##EQU1##

In Vivo Antibacterial Evaluation (Table 3)

The therapeutic effects of tetracyclines are determined against acutelethal infections with various staphylococcal and E. coli strains.Female mice, strain CD-1 Charles River Laboratories, (20±2 grams) arechallenged by an intraperitoneal injection of sufficient bacteria(suspended in broth or hog gastric mucin) to kill non-treated controlswithin 24-48 hours. Antibacterial agents, contained in 0.5 ml of 0.2%aqueous agar, are administered subcutaneously or orally 30 minutes afterinfection. When an oral dosing schedule is used, animals are deprived cffood for 5 hours before and 2 hours after infection. Five mice aretreated at each dose level. The 7 day survival ratios from threeseparate tests are pooled for calculation of median effective dose(ED₅₀).

Legend for Tables I-III

A=9-Amino-7-(dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride

B=7-(Dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride(minocycline hydrochloride)

C=9-Amino-7-(diethylamino)-6-demethyl-6-deoxytetracycline sulfate

D=7-(Diethylamino)-6-demethyl-6-deoxytetracycline sulfate

                                      TABLE 1                                     __________________________________________________________________________    In Vitro Antibacterial Activity of 6-Demethyl-6-deoxytetracycline             Derivatives                                                                                         MIC (μg/ml)                                          Organism*             A.sup.a                                                                            B    C.sup.a                                                                           D                                         __________________________________________________________________________    S. aureus UBMS 88-5 (tetM)                                                                          0.06 4    0.5 16                                        S. aureus UBMS 88-4 (tetracycline-sensitive)                                                        0.015                                                                              0.008                                                                              0.03                                                                              0.03                                      S. aureus UBMS 90-1 (tetM)                                                                          0.25 4    2   8                                         S. aureus UBMS 90-2 (tetM)                                                                          0.06 1    0.25                                                                              8                                         S. aureus UBMS 90-3 (tetracycline-sensitive)                                                        0.015                                                                              0.015                                                                              0.03                                                                              0.03                                      S. aureus UBMS 88-7 (tetK)                                                                          0.12 0.03 0.06                                                                              0.12                                      S. aureus IVES 2943 (methicillin-resistant)                                                         0.5  1    0.25                                                                              8                                         S. aureus IVES 1983 (methicillin-resistant)                                                         0.5  1    0.25                                                                              8                                         S. aureus CI 2371 (methicillin-resistant)                                                           0.5  4    NA  NA                                        S. aureus CI3300 (methicillin-resistant)                                                            0.25 8    NA  NA                                        Coagulase negative staphylococci CI 664                                                             0.003                                                                              0.015                                                                              NA  NA                                        Coagulase negative staphylococci CI 535                                                             1    8    NA  NA                                        S. haemolyticus AVAH 88-3                                                                           0.06 0.12 0.12                                                                              NA                                        E. faecalis AMV 120 (tetM)                                                                          4    16   NA  NA                                        E. faecalis PAM 211 (tetN)                                                                          4    16   NA  NA                                        E. faecalis 12201 (vancomycin-resistant)                                                            0.5  4    NA  NA                                        E. faecalis CI 2735   0.5  4    NA  NA                                        E. coli UBMS 88-1 (tetB)                                                                            >32  8    2   32                                        E. coli UBMS 88-2 (tetracycline-sensitive)                                                          0.25 0.5  0.5 2                                         E. coli UBMS 89-1 (tetM)                                                                            1    8    2   NA                                        E. coli UBMS 89-2 (tetracycline-sensitive)                                                          0.5  0.5  0.5 4                                         E. coli UBMS 90-4 (tetM)                                                                            4    >32  32  >32                                       E. coli UBMS 90-5 (tetracycline-sensitive)                                                          0.25 0.5  1   2                                         M. morganii NEMC 87-119                                                                             2    2    2   32                                        S. marcescens FPOR 87-33                                                                            4    2    4   32                                        P. aeruginosa ATCC 27853                                                                            2    4    8   32                                        X. maltophilia FPOR 87-210                                                                          0.12 0.06 0.12                                                                              0.25                                      E. coli ATCC25922     0.25 0.25 0.50                                                                              1                                         E. faecalis ATCC 29212                                                                              0.06 0.25 0.12                                                                              8                                         S. aureus ATCC 29213  0.008                                                                              ≦0.004                                                                      ≦0.015                                                                     <0.015                                    __________________________________________________________________________     .sup.a In vitro assay is done in the presence of cysteine (0.05%).            Antibacterial potency of B and D was not enhanced in the presence of          cysteine.                                                                     *The tetM resistance determinant protects ribosomes from tetracycline, th     tetK determinant promotes efflux of the drug from the cell.              

                                      TABLE 2                                     __________________________________________________________________________    In Vitro Protein Translation with E. coli S30 Ribosomes                       __________________________________________________________________________    Wild Type Version                                                                      B                          A                                                  Wild Type S30 no DTT*                                                                      Wild Type S30 with DTT*                                                                     Wild Type S30 no DTT*                                                                       Wild Type S30 with DTT      Reaction Counts                                                                             % Inhibition                                                                          Counts                                                                              % Inhibition                                                                          Counts                                                                              % Inhibition                                                                          Counts                                                                             %                      __________________________________________________________________________                                                           Inhibition             Control DMSO                                                                           457268       872132        457268        872132                      Plus compound                                                                 1.0 mg/ml                                                                               57696                                                                             87       58885                                                                              93       52595                                                                              88       38121                                                                             96                     1:2 Dilution                                                                            76804                                                                             83       78961                                                                              91       80494                                                                              82       60891                                                                             93                     1:4 Dilution                                                                            53400                                                                             88      111971                                                                              87       95015                                                                              79       92203                                                                             89                     1:8 Dilution                                                                           162405                                                                             64      149792                                                                              83      130209                                                                              72      143215                                                                             84                     1:16 Dilution                                                                          213077                                                                             53      207484                                                                              76      205392                                                                              55      214831                                                                             75                     1:32 Dilution                                                                          306650                                                                             33      289304                                                                              67      297786                                                                              35      238321                                                                             73                     1:64 Dilution                                                                          457601                                                                             0       518601                                                                              41      245494                                                                              46      380103                                                                             56                     tetM Variant                                                                           tetM S30 no DTT*                                                                           tetM S30 with DTT*                                                                          tetM S30 no DTT*                                                                             tetM S30 with DTT*         Reaction Counts                                                                             % Inhibition                                                                          Counts                                                                              % Inhibition                                                                          Counts                                                                              % Inhibition                                                                          Counts                                                                             %                      __________________________________________________________________________                                                           Inhibition             Control DMSO                                                                           247938       447247        247938        447247                      Plus compound                                                                 1.0 mg/ml                                                                              281528                                                                             0       371475                                                                              17      129780                                                                              48      145068                                                                             68                     1:2 Dilution                                                                           258633                                                                             0       331439                                                                              26      158861                                                                              36      168574                                                                             62                     1:4 Dilution                                                                           248904                                                                             0       373168                                                                              17      203184                                                                              18      226708                                                                             49                     1:8 Dilution                                                                           289595                                                                             0       421533                                                                               6      231447                                                                               7      284606                                                                             36                     1:16 Dilution                                                                          287498                                                                             0       493679                                                                               0      305633                                                                               0      375989                                                                             16                     1:32 Dilution                                                                          262329                                                                             0       490283                                                                               0      349994                                                                               0      412967                                                                              8                     1:64 Dilution                                                                          249242                                                                             0       452837                                                                               0      310723                                                                               0      507461                                                                              0                     __________________________________________________________________________     *Dithiothreitol (DTT) is used as an oxygen scavenger.                    

                  TABLE 3                                                         ______________________________________                                        Effects of Compounds A and B on Acute Lethal                                  Infections in Mice                                                                          Route                                                                         of Antibiotic                                                                           ED.sub.50 (mg/kg).sup.+                               Organism        Administration*                                                                           A      B                                          ______________________________________                                        E. coli 311 (sens)                                                                            Subcutaneous                                                                              2.8    3.1                                        E. coli UBMS 90-4 (tetM)                                                                      Subcutaneous                                                                              24     >256                                       S. aureus UBMS 90-1 (tetM)                                                                    Subcutaneous                                                                              0.30   1.7                                        S. aureus UBMS 90-2 (tetM)                                                                    Oral        1.6    1.8                                                        Subcutaneous                                                                              0.53   1.8                                        S. aureus Smith (sens)                                                                        Oral        0.81   0.53                                                       Subcutaneous                                                                              0.34   0.32                                       ______________________________________                                         *Single Dose                                                                  .sup.+ Median effective dose protecting 50% of the infected mice         

Testing Results

As seen from the above testing, the compounds according to the presentinvention display good activity against a spectrum of tetracyclinesensitive and resistant Gram-positive and Gram-negative bacteria,especially strains of E. coli, S. aureus and E. faecalis containing thetetM or tetK resistant determinants. As illustrated in Table I,9-amino-7-(dimethylamino) 6-demthyl-6-deoxytetracycline hydrochloride(A) shows good in vitro activity against tetracycline resistant strainscarrying the tetM resistance determinant such as S. aureus UBMS 88-5,S-aureus UBMS 90-1 and 90-2, E. coli UBMS 89-1 and 90-4; and is equallyas effective as 7-(dimethylamino)-6-demethyl-6-deoxytetracyclinehydrochloride (B) vs. susceptible strains.7-(dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride (B) and9-amino-7-(dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride(A) are assayed in vitro for their ability to inhibit protein synthesis,taking place on either wild type or tetM protected ribosomes, using acoupled transcription and translation system. Similarly,9-amino-7-(diethylamino)-6-demethyl -6-deoxytetracycline sulfate (C)shows enhancement of antibacterial activity versus 7-(diethylamino)-6-demethyl-6-deoxytetracycline sulfate (D).

Both compounds (A & B) are found to effectively inhibit proteinsynthesis on wild type ribosomes, having equivalent levels of activity(Table II). 7-(dimethylamino)-6-demethyl-6-deoxytetracyclinehydrochloride (B) is unable to inhibit protein synthesis occurring ontetM protected ribosomes. In contrast,9-amino-7-(dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride(A) is effective in inhibiting protein synthesis occurring on tetMprotected ribosomes, although higher drug levels are required to achievesimilar levels of inhibition relative to wild type ribosomes.

The enhanced activity of9-amino-7-(dimethylamino)-6-demethyl-6-deoxytetracycline sulfate (A)against tetracycline susceptible and resistant organisms (tetM) isdemonstrated in Table 3 for animals infected with representativebacteria Lowered ED₅₀ 's are obtained with9-amino-7-(dimethylamino)-6-demethyl-6-deoxytetracycline sulfate (A)than with 7-(dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride(B) in mice with of S. aureus and E. coli which carry the tetMresistance determinant. Similar ED₅₀ 's are obtained with9-amino-7-(dimethylamino)-6-demethyl-6-deoxytetracycline sulfate (A) and7-(dimethylamino)-6-demethyl-6-deoxytetracycline hydrochloride (B)against infections with minocycline susceptible organisms.

As can be seen from Tables 1 and 3, the new 9-amino-7-(substitutedamino)-6-demethyl-6-deoxytetracyclines may be used to prevent or controlimportant veterinary diseases such as mastitis, diarrhea, urinary tractinfections, skin infections, ear infections, wound infections and thelike.

The improved efficacy of the new 9-amino-7(substitutedamino)-6-demethyl-6-deoxytetracyclines is evidenced by the in vitroactivity against isogenic strains into which the resistant determinants,such as tetM, were cloned (Table 1); the inhibition of protein synthesisby tetM resistant ribosomes (Table 2); and the in vivo activity againstexperimental infections caused by strains resistant to thetetracyclines, due to the presence of resistant determinants, tetM(Table 3).

When the compounds are employed as antibacterials, they can be combinedwith one or more pharmaceutically acceptable carriers, for example,solvents, diluents and the like, and may be administered orally in suchforms as tablets, capsules, dispersible powders, granules, orsuspensions containing, for example, from about 0.05 to 5% of suspendingagent, syrups containing, for example, from about 10 to 50% of sugar,and elixirs containing, for example, from about 20 to 50% ethanol, andthe like, or parenterally in the form of sterile injectable solutions orsuspensions containing from about 0.05 to 5% suspending agent in anisotonic medium. Such pharmaceutical preparations may contain, forexample, from about 25 to about 90% of the active ingredient incombination with the carrier, more usually between about 5% and 60% byweight.

An effective amount of compound from 2.0 mg/kg of body weight to 100.0mg/kg of body weight should be administered one to four times per dayvia any typical route of administration including but not limited tooral, parenteral (including subcutaneous, intravenous, intramuscular,intrasternal injection or infusion techniques), topical or rectal, indosage unit formulations containing conventional non-toxicpharmaceutically acceptable carriers, adjuvants and vehicles. It will beunderstood, however, that the specific dose level and frequency ofdosage for any particular patient may be varied and will depend upon avariety of factors including the activity of the specific compoundemployed, the metabolic stability and length of action of that compound,the age, body weight, general health, sex, diet, mode and time ofadministration, rate of excretion, drug combination, the severity of theparticular condition, and the host undergoing therapy.

These active compounds may be administered orally as well as byintravenous, intramuscular, or subcutaneous routes. Solid carriersinclude starch, lactose, dicalcium phosphate, microcrystallinecellulose, sucrose and kaolin, while liquid carriers include sterilewater, polyethylene glycols, non-ionic surfactants and edible oils suchas corn, peanut and sesame oils, as are appropriate to the nature of theactive ingredient and the particular form of administration desired.Adjuvants customarily employed in the preparation of pharmaceuticalcompositions may be advantageously included, such as flavoring agents,coloring agents, preserving agents, and antioxidants, for example,vitamin E, ascorbic acid, BHT and BHA.

The preferred pharmaceutical compositions from the standpoint of ease ofpreparation and administration are solid compositions, particularlytablets and hard-filled or liquid-filled capsules. Oral administrationof the compounds is preferred.

These active compounds may also be administered parenterally orintraperitoneally. Solutions or suspensions of these active compounds asa free base or pharmacologically acceptable salt can be prepared inwater suitably mixed with a surfactant such as hydroxypropylcellulose.Dispersions can also be prepared in glycerol, liquid polyethyleneglycols and mixtures thereof in oils. Under ordinary conditions ofstorage and use, these preparations contain a preservative to preventthe growth of microorganisms.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. In all cases, the form must be sterile and must be fluid tothe extent that easy syringability exists. It must be stable under theconditions of manufacture and storage and must be preserved against thecontaminating action of microorganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (e.g., glycerol, propylene glycol and liquidpolyethylene glycol), suitable mixtures thereof, and vegetable oil.

The invention will be more fully described in conjunction with thefollowing specific examples which are not to be construed as limitingthe scope of the invention.

EXAMPLE 1 [4S-(4α,12aα)]-4,7-Bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate (1:1)

To a stirred ice bath cooled solution of 0.444 g of[4S-(4α,12aα)]-4,7-bis(dimethylamino)-1,4,-4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidehydrochloride, prepared by the procedure described in U.S. Pat. No.3,226,436, dissolved in 15 ml of sulfuric acid is added 0.101 g ofsodium nitrate. The mixture is stirred in the cold for 45 minutesfollowed by the dropwise addition to 500 ml of diethyl ether. Theresulting solid is collected, washed with diethyl ether and dried togive 0.6 g of the desired product as a solid.

MS(FAB): m/z 503(M+H) and 601(M+H₂ SO₄ +H).

EXAMPLE 2[4S-(4α,12aα)]-7-(Diethylamino)-4-dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate (1:2)

To a stirred ice cooled solution of 0.660 g of[4S-(4α,12aα)]-7-(diethylamino)-4-(dimethylamino)1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide hydrochloride, prepared by theprocedure described in U.S. Pat. No. 3,226,436, dissolved in 15 ml ofsulfuric acid is added 0.151 g of sodium nitrate. The mixture is stirredin the cold followed by dropwise addition to 500 ml of diethyl ether.The resulting solid is collected, washed with diethyl ether and dried togive 0.8 g of the desired product as a solid.

MS(FAB): m/z 531(M+H) and 629(M+H₂ SO₄ +H).

EXAMPLE 3

[4S-(4α,12aα)]-9-Amino-4,7-bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate (1:1)

A mixture of 2.0 g of product from Example 1 in 20 ml of2-methoxyethanol is stirred for 10 minutes and filtered. The filtrate isshaken, in a pressure bottle, with 1.0 g of 10% palladium-on-carbon and5 ml of 2N sulfuric acid, under 30 lbs. of hydrogen pressure, for 1hour. The reaction is filtered and the filtrate concentrated in vacuo tohalf volume. The solution is poured into 100 ml of diethyl ether, thesolid collected, washed with diethyl ether and dried to give 1.6 g ofthe desired product as a solid.

MS(FAB): m/z 473(M+H).

EXAMPLE 4[4S-(4α,12aα)]-9-Amino-4,7-bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12α-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide hydrochloride (1:1)

A mixture of 20.0 g of product from Example 1 in 250 ml of2-methoxyethanol is stirred for 10 minutes and filtered. The filtrate isshaken, in a pressure bottle, with 10.0 g of 10% palladium-on-carbon and100 ml of 1N ethanolic hydrogen chloride, under 30 lbs. of hydrogenpressure, for 1 hour. The reaction mixture is filtered and the filtrateconcentrated in vacuo to half volume. The solution is poured into 1 L ofdiethyl ether, the solid collected, washed with diethyl ether and driedto give 16.0 g of the desired product as an oil. The oil is suspended in20 ml of distilled water, made acidic with 2.8 ml of 32% hydrochloricacid and decolorized with charcoal. The mixture is filtered throughdiatomaceous earth and made basic (pH 4.0) with concentrated ammoniumhydroxide. The solid is collected at 4° C., washed with pH 4 water anddried in vacuo to give 14.2 g of the desired product as a solid.

¹ H NMR (CD₃ SOCD₃): δ4.19(s,1H,4-H) and 7.29(s,1H,8-H).

MS(FAB): m/z 473(M+H).

Analysis for C₂₃ H₂₈ N₄ O₇ HCl 6.7% H₂ O: Calc'd: C,50.64; H,6.10;N,10.27; Cl,6.50: Found: C,50.72; H,6.07; N,10.27; Cl,6.62.

EXAMPLE 5 [4S-(4α,12aα)]-Amino-4,7-bis(dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide p-toluenesulfonate(1:1)

The title compound is prepared by the procedure of Example 4, using 20 gof product from Example 1, to give 16.0 g of the desired product as thefree base. The free base is suspended in 20 ml of distilled water, madeacidic with p-toluenesulfonic acid monohydrate and decolorized withcharcoal. The mixture is filtered through diatomaceous earth and madebasic (pH 4.0) with concentrated ammonium hydroxide. The solid iscollected at 4° C., washed with pH 4 water and dried in vacuo to give16.0 g of the desired product.

EXAMPLE 6 [4S-(4α,12aα)]-9-Amino-7-(diethylamino)-4-dimethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate (1:2)

The title compound is prepared by the procedure of Example 3, using 2.1g of product from Example 2, to give 1.5 g of the desired product as asolid.

¹ H NMR (CD₃ SOCD₃): δ4.25(s,1H,4-H) and 7.27(s,1H,8-H).

MS(FAB): m/z 501(M+H) and 599(M+H₂ SO₄ +H).

EXAMPLE 7 [4S-(4α,12aα)]-4-(Dimethylamino)-7-(ethylmethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12atetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide hydrochloride (1:1)

A solution of 0.460 g of[4S-(4α,12aα)]-4-(dimethylamino)-7-(ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidehydrochloride, prepared by the procedure described in U.S. Pat. No.3,226,436, 0.5 ml of 97% formic acid and 0.75 ml of 40% aqueousformaldehyde is heated at reflux temperature for 2 hours. The reactionmixture is cooled, concentrated in vacuo to half volume and poured intodiethyl ether. The resulting solid is collected, washed with diethylether and dried to give 0.3 g of the desired product as a solid.

EXAMPLE 8 [4S-(4α,12aα)]-4-(Dimethylamino)-7-(ethylmethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydro-9-nitro-1.11-dioxo-2-naphthacenecarboxamide sulfate (1:1)

The title compound is prepared by the procedure of Example 1, using 0.46g of product from Example 7 to give 0.5 g of the desired product as asolid.

EXAMPLE 9 [4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(ethylmethylamino)-1,4,4a,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate (1:1)

The title compound is prepared by the procedure of Example 3, using 1.0g of product from Example 8, to give 0.8 g of the desired product as asolid.

EXAMPLE 10[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate (1:1)

The title compound is prepared by the procedure of Example 1, using 0.48g of[4S-(4α,12aα)]-4-(dimethylamino)-7-[(1-methylethyl)amino]-1,4,4a,5,5a,-6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidehydrochloride, prepared by the procedure described in U.S. Pat. No.3,226,436, to give 0.5 g of the desired product as a solid.

EXAMPLE 11[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylethyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12atetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate (1:1)

The title compound is prepared by the procedure of Example 3, using 2.1g of product from Example 10, to give 1.5 g of the desired product as asolid.

EXAMPLE 12[4S-(4α,12aα)]-4-(Dimethylamino)-7-(ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,1012,12a -tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate(1:1)

The title compound is prepared by the procedure of Example 1, using 0.96g of [4S-(4α,12aα)]-4-dimethylamino)-7-(ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamideprepared by the procedure described in U.S. Pat. No. 3,226,436, to give0.9 g of the desired product as a solid.

EXAMPLE 13 [4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate (1:1)

The title compound is prepared by the procedure of Example 3, using 1.0g of product from Example 12, to give 0.7 g of the desired product as asolid.

EXAMPLES 14-35

Substantially following the methods described in detail hereinabove inExamples 3 and 9, the compounds of this invention listed below inExamples 14-35 are prepared.

EXAMPLE 14

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(methyl-propylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 15

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(butylmethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 16

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[1-methylpropylamino)methylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 17

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(2-methylpropyl)methylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 18

[4S-(4α,12aα))]-9-Amino-4-(dimethylamino)-7-(ethylpropylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 19

[4S-(4α,12aα))]-9-Amino-4-(dimethylamino)-7-[(1-methylethyl)ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 20

[4S-(4α,12aα) )]-9-Amino-4-(dimethylamino)-7-(butylethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 21

[4S-(4α,12aα))]-9-Amino-4-(dimethylamino)-7-[(1-methylpropyl)ethylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 22

[4S-(4α,12aα))]-9-Amino-4-(dimethylamino)-7-[(2-methylpropyl)ethylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 23

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(dipropylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 24

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methyethyl)propylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 25

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(butylpropylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 26

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylpropyl)propylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 27

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(2-methylpropyl)propylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11=dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 28

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylethyl)butylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 29

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylethyl)(1-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 30

4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylethyl)(2-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10, 12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 31

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(dibutylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 32

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylpropyl)butylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 33

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(2-methylpropyl)butylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 34

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 35

4S-(4α,12aα)]-9Amino-4-(dimethylamino)-7-[(1-methylpropyl)(2-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLES 36-39

Substantially following the methods described in detail hereinabove inExamples 3 and 11, the compounds of this invention listed below inExamples 36-39 are prepared.

EXAMPLE 36

[4S-(4,60 ,12aα)]-9-Amino-4-(dimethylamino)-7-(propylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 37

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-(butylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 38

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(2-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 39

[4S-(4α,12aα)]-9-Amino-4-(dimethylamino)-7-[(1,1-dimethylethyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLES 40-65

Substantially following the methods described in detail hereinabove inExamples 1 and 2, the compounds of this invention listed below inExamples 40-65 are prepared.

EXAMPLE 40

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(methylpropylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 41

4S-(4α,12aα)]-4-(Dimethylamino)-7-(butylmethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 42

4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylpropylamino)methylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 43

4S-(4α,12aα)]-4-(Dimethylamino)-7-[(2-methylpropyl)methylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2naphthacenecarboxamidesulfate.

EXAMPLE 44

4S-(4α,12aα)]-4-(Dimethylamino)-7-(ethylpropylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 45

4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)ethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2naphthacenecarboxamidesulfate

EXAMPLE 46

[4S-4α,12aα)]-4-(Dimethylamino)-7-(butylethylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 47

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylpropyl)ethylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 48

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(2-methylpropyl)ethylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 49

[4S-(4α,12aα)1-4-{Dimethylamino)-7-(dipropylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 50

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)propylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2naphthacenecarboxamidesulfate.

EXAMPLE 51

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(butylpropylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthaoeneoarboxamide sulfate.

EXAMPLE 52

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylpropyl)propylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2naphthacenecarboxamidesulfate.

EXAMPLE 53

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(2-methylpropyl)propylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2naphthacenecarboxamidesulfate.

EXAMPLE 54

4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)butylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 55

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)(1-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2naphthacenecarboxamidesulfate.

EXAMPLE 56

[4S-4α,12aα)]-4-(Dimethylamino)-7-[(1-methylethyl)(2-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2naphthacenecarboxamidesulfate.

EXAMPLE 57

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(dibutylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 58

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylpropyl)butylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2naphthacenecarboxamidesulfate.

EXAMPLE 59

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(2-methylpropyl)butylamino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2naphthacenecarboxamidesulfate.

EXAMPLE 60

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2naphthacenecarboxamidesulfate.

EXAMPLE 61

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1-methylpropyl)(2-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamidesulfate.

EXAMPLE 62

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(propylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 63

[4S-(4α,12aα)]-4-(Dimethylamino)-7-(butylamino)-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 64

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(2-methylpropyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate.

EXAMPLE 65

[4S-(4α,12aα)]-4-(Dimethylamino)-7-[(1,1-dimethyethyl)amino]-1,4,4a,5,5a,6,11,12a-octahydro-3,10,12,12a-tetrahydroxy-9-nitro-1,11-dioxo-2-naphthacenecarboxamide sulfate.

We claim:
 1. A compound of the formula: ##STR16## wherein: R=NR₁ R₂,andwhen R₁ =hydrogen, R₂ =methyl, ethyl, n-propyl, 1-methylethyl, n-butyl,1-methylpropyl, 2-methylpropyl or 1,1-dimethylethyl; and when R₁ =methylor ethyl, R₂ =methyl, ethyl, n-propyl, 1-methylethyl, n-butyl,1-methylpropyl or 2-methylpropyl; and when R₁ =n-propyl, R₂ =n-propyl,1-methylethyl, n-butyl, 1-methylpropyl or 2-methylpropyl; and when R₁=1-methylethyl, R₂ =n-butyl, 1-methylpropyl or 2-methylpropyl; and whenR₁ =n-butyl, R₂ =n-butyl, 1-methylpropyl or 2-methylpropyl; and when R₁=1-methylethyl, R₂ =2-methylpropyl; R₃ is selected from (C₇ -C₉)aralkylgroup; a heterocycle group selected from 2 or 3-furanyl, 2 or 3-thienyl,2,3 or 4-pyridyl, di(C₁ -C₃)alkyl substituted pyridyl, benzofuranyl,benzothienyl, quinolinyl; R₄ is selected from hydrogen; straight orbranched (C₁ -C₃)alkyl group; (C₆ -C₁₀)aryl group; (C₇ -C₉)aralkylgroup; a heterocycle group selected from 2 or 3-furanyl, 2 or 3-thienyl,2,3 or 4-pyridyl, di(C₁ -C₃)-alkyl substituted pyridyl, benzofuranyl,benzothienyl, quinolinyl or --(CH₂)_(n) when n=1-4 and R₆ is selectedfrom hydrogen; straight or branched (C₁ -C₃)alkyl; or (C₆ -C₁₀)aryl; orR₃ and R₄ taken together are--(CH₂)₂ W(CH₂)--₂, wherein W is selectedfrom (CH₂)_(n) and n=0-1, --NH, --N(C₁ -C₃)alkyl, --N(C₁ -C₄)alkoxy,oxygen, sulfur or substituted congeners selected from (L or D) proline,ethyl (L or D) prolinate, morpholine, pyrrolidine or piperidine; and thepharmacologically acceptable organic and inorganic salts and metalcomplexes.
 2. The compound according to claim 1, wherein:R=NR₁ R₂, andwhen R₁ =hydrogen, R₂ =ethyl, n-propyl, 1-methylethyl, n-butyl,1-methylpropyl, 2-methylpropyl or 1,1-dimethylethyl; and when R₁=methyl, R₂ =methyl, ethyl, n-propyl, n-butyl, 1-methylpropyl or2-methylpropyl; and when R₁ =ethyl, R₂ =ethyl, n-propyl, n-butyl, or2-methylpropyl; and when R₁ =n-propyl, R₂ =n-propyl, n-butyl or2-methylpropyl; and when R₁ =n-butyl, R₂ =n-butyl or 2-methylpropyl; R₃is selected from (C₇ -C₉)aralkyl group; a heterocycle group selectedfrom 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkylsubstituted pyridyl, benzofuranyl, benzothienyl, quinolinyl or (CH₂)_(n)COOR₅ when n=1-4 and R₅ is selected from hydrogen; straight or branched(C₁ -C₃)alkyl group; or (C₆ -C₁₀)aryl group; R₄ is selected fromstraight or branched (C₁ -C₃)alkyl group; (C₆ -C₁₀)aryl group; (C₇-C₉)aralkyl group; a heterocycle group selected from 2 or 3-furanyl, 2or 3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)alkyl substituted pyridyl,benzofuranyl, benzothienyl, quinolinyl or--(CH₂)_(n) COOR₆ when n=1-4and R₆ is selected from hydrogen; straight or branched (C₁ -C₃)alkyl; or(C₆ -C₁₀)-aryl; or R₃ and R₄ taken together are --(CH₂)₂ W(CH₂)--₂,wherein W is selected from (CH₂)_(n) and n=0-1, --NH, --N(C₁ -C₃)alkyl,--N(C₁ -C₄)alkoxy, oxygen, sulfur or substituted congeners selected from(L or D) proline or ethyl (L or D) prolinate, morpholine, pyrrolidine orpiperidine; and the pharmacologically acceptable organic and inorganicsalts and metal complexes.
 3. The compound according to claim 1,wherein:R=NR₁ R₂, and when R₁ =hydrogen, R₂ =ethyl, n-propyl or1-methylethyl; and when R₁ =methyl; R₂ =methyl, ethyl or n-propyl; andwhen R₁ =ethyl; R₂ =ethyl; R₃ and R₄ taken together are --(CH₂)₂W(CH₂)--₂, wherein W is selected from (CH₂)_(n) and n=0-1, --NH, --N(C₁-C₃)alkyl, --N(C₁ -C₄)alkoxy, oxygen, sulfur or substituted congenersselected from (L or D) proline or ethyl (L or D) prolinate, morpholine,pyrrolidine or piperidine; and the pharmacologically acceptable organicand inorganic salts and metal complexes.
 4. A compound of the formula:##STR17## wherein: R=NR₁ R₂,and when R₁ =hydrogen, R₂ =methyl, ethyl,n-propyl, 1 -methylethyl, n-butyl, 1-methylpropyl, 2-methylpropyl or1,1-dimethylethyl; and when R₁ =methyl or ethyl, R₂ =methyl, ethyl,n-propyl, 1-methylethyl, n-butyl, 1-methylpropyl or 2-methylpropyl; andwhen R₁ =n-propyl, R₂ =n-propyl, 1-methylethyl, n-butyl, 1-methylpropylor 2-methylpropyl; and when R₁ =1-methylethyl, R₂ =n-butyl,1-methylpropyl or 2-methylpropyl; and when R₁ =n-butyl, R₂ =n-butyl,1-methylpropyl or 2-methylpropyl; and when R₁ =1-methylpropyl, R₂=2-methylpropyl; R₃ is selected from a heterocycle group selected from 2or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃)-alkylsubstituted pyridyl, benzofuranyl, benzothienyl, quinolinyl; R₄ isselected from hydrogen; straight or branched (C₁ -C₃)alkyl group; (C₆-C₁₀)aryl group; (C₇ -C₉)aralkyl group; a heterocycle group selectedfrom 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or 4-pyridyl, di(C₁ -C₃) -alkylsubstituted pyridyl, benzofuranyl, benzothienyl, quinolinyl or--(CH₂)_(n) COOR₆ when n=1-4 and R₆ is selected from hydrogen; straightor branched (C₁ -C₃)alkyl; or (C₆ -C₁₀)aryl; or R₃ and R₄ taken togetherare --(CH₂)₂ W(CH₂)--₂, wherein W is selected from (CH₂)_(n) and n=0-1,--NH, --N(C₁ -C₃)alkyl, --N(C₁ -C₄)alkoxy, oxygen, sulfur or substitutedcongeners selected from (L or D) proline, ethyl (L or D) prolinate,morpholine, pyrrolidine or piperidine; and the pharmacologicallyacceptable organic and inorganic salts and metal complexes.
 5. Thecompound according to claim 4, whereinR=NR₁ R₂, and when R₁ =hydrogen,R₂ =ethyl, n-propyl, 1-methylethyl, n-butyl, 1-methylpropyl,2-methylpropyl or 1,1-dimethylethyl: and when R₁ =methyl, R₂ =methyl,ethyl, n-propyl, n-butyl, 1-methylpropyl, or 2-methylpropyl; and when R₁=ethyl, R₂ =ethyl, n-propyl, n-butyl, or 2-methylpropyl; and when R₁=n-propyl, R₂ =n-propyl, n-butyl, or 2-methylpropyl; and when R₁=n-butyl, R₂ =n-butyl or 2-methylpropyl; R₃ is selected from aheterocycle group selected from 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or4-pyridyl, di(C₁ -C₃)alkyl substituted pyridyl, benzofuranyl,benzothienyl, quinolinyl; R₄ is selected from straight or branched (C₁-C₃)alkyl group; (C₆ -C₁₀)aryl group; (C₇ -C₉)aralkyl group; aheterocycle group selected from 2 or 3-furanyl, 2 or 3-thienyl, 2,3 or4-pyridyl, di(C₁ -C₃)alkyl substituted pyridyl, benzofuranyl,benzothienyl, quinolinyl or --(CH₂)_(n) COOR₆ when n=1-4 and R₆ isselected from hydrogen; straight or branched (C₁ -C₃)alkyl; or (C₆-C₁₀)aryl; or R₃ and R₄ taken together are --(CH₂)₂ W(CH₂)--₂, wherein Wis selected from (CH₂)_(n) and n=0-1, --NH, --N(C₁ -C₃)alkyl, --N(C₁-C₄)alkoxy, oxygen sulfur or substituted congeners selected from (L orD) proline or ethyl (L or D) prolinate, morpholine, pyrrolidine orpiperidine; and the pharmacologically acceptable organic and inorganicsalts and metal complexes.
 6. The compound according to claim 4,wherein:R=NR₁ R₂, and when R₁ =hydrogen, R₂ =ethyl, n-propyl or1-methylethyl; and when R₁ =methyl, R₂ =methyl, ethyl or n-propyl; andwhen R₁ =ethyl, R₂ =ethyl; R₃ and R₄ taken together are --(CH₂)₂W(CH₂)--₂, wherein W is selected from (CH₂)_(n) and n=0-1, --NH, --(C₁-C₃)alkyl, --N(C_(1-C) ₄)alkoxy, oxygen, sulfur or substitute congenersselected from (L or D) proline or ethyl (L or D) prolinate, morpholine,pyrrolidine or piperidine; and the pharmacologically acceptable organicand inorganic salts and metal complexes.
 7. The compound according toclaim 1 wherein said inorganic salts comprise: hydrochloric,hydrobromic, hydroiodic, phosphoric, nitric or sulfate.
 8. The compoundaccording to claim 1 wherein said organic salts comprise: acetate,benzoate, citrate, cysteine or other amino acids, fumarate, glycolate,maleate, succinate, tartrate, alkylsulfonate or arylsulfonate.
 9. Thecompound according to claim 1 wherein said metal complexes comprise:aluminum, calcium, iron, magnesium, manganese and complex salts.
 10. Thecompound according to claim 4 wherein said inorganic salts comprise:hydrochloric, hydrobromic, hydroiodic, phosphoric, nitric or sulfate.11. The compound according to claim 4 wherein said organic saltscomprise: acetate, benzoate, citrate, cysteine or other amino acids,fumarate, glycolate, maleate, succinate, tartrate, alkylsulfonate orarylsulfonate.
 12. The compound according to claim 4 wherein said metalcomplexes comprise: aluminum, calcium, iron, magnesium, manganese andcomplex salts.
 13. A method for the prevention, treatment or control ofbacterial infections in warm-blooded animals which comprisesadministering to said animal a pharmacologically effective amount of acompound according to claim
 1. 14. A pharmaceutical composition ofmatter comprising a compound according to claim 1 in association with apharmaceutically acceptable carrier.
 15. A veterinary composition whichcomprises a pharmacologically effective amount of a compound accordingto claim 1 and pharmaceutically acceptable carrier.
 16. A method for theprevention, treatment or control of bacterial infections in warm-bloodedanimals caused by bacteria having the TetM and TetK resistantdeterminants which comprises administering to said animal apharmacologically effective amount of a compound according to claim 1.17. An antibacterial pharmaceutical composition of matter comprising aneffective amount of a compound according to claim 1 and an effectiveamount of an antioxidant in association with a pharmaceuticallyacceptable carrier.
 18. The composition of claim 17 wherein saidantioxidant comprises: cysteine, cysteine hydrochloride, dithiothreitol,ascorbic acid, monothioglycerol or thioglycolic acid.